Division of Nutrition, St. John's Research Institute, A recognized research centre of University of Mysore, Bangalore, Karnataka, India.
Department of Pathology, St John's Medical College Hospital, Bangalore, Karnataka, India.
Eur J Clin Nutr. 2022 May;76(5):722-729. doi: 10.1038/s41430-021-01003-3. Epub 2021 Sep 10.
BACKGROUND/OBJECTIVES: The current study aimed to identify suitable reference miRNA for placental miRNA expression analysis in a set of well-characterized and fetal-sex balanced small- (SGA) and appropriate- (AGA) for gestational age full-term singleton pregnancies.
SUBJECTS/METHODS: In this retrospective study, placental samples (n = 106) from 35 SGA (19 male and 16 female) and 71 AGA (30 male and 41 female) full-term singleton pregnancies were utilized. Placental transcript abundance of three widely used reference miRNAs [miR-16-5p and Small nucleolar RNAs (snoRNAs) RNU44 and RNU48] were assessed by real-time quantitative PCR. Raw cycle threshold (C) analysis and RefFinder tool analysis were conducted for evaluating stability of expression of these miRNAs.
Raw C values of miR-16-5p were similar between SGA and AGA births (P = 0.140) and between male and female births within SGA (P = 0.159) and AGA (P = 0.060) births while that of RNU44 and RNU48 were higher in SGA births (P = 0.008 and 0.006 respectively) and in male births within the SGA group (P = 0.005) for RNU44 and in female births within the AGA group (P = 0.048) for RNU48. Across all 106 samples tested using the RefFinder tool, miR-16-5p and RNU44 were equally stable reference miRNAs.
We recommend miR-16-5p and RNU44 as suitable reference miRNAs for placental samples from settings similar to our study.
背景/目的:本研究旨在鉴定一组特征明确且胎儿性别均衡的小(SGA)和适当(AGA)胎龄足月单胎妊娠中胎盘 miRNA 表达分析的合适参考 miRNA。
受试者/方法:在这项回顾性研究中,使用了 35 例 SGA(19 名男性和 16 名女性)和 71 例 AGA(30 名男性和 41 名女性)足月单胎妊娠的胎盘样本。通过实时定量 PCR 评估三种广泛使用的参考 miRNA [miR-16-5p 和小核仁 RNA(snoRNA)RNU44 和 RNU48]的胎盘转录丰度。进行了原始循环阈值(C)分析和 RefFinder 工具分析,以评估这些 miRNA 表达稳定性。
SGA 和 AGA 出生之间的 miR-16-5p 的原始 C 值相似(P=0.140),SGA 和 AGA 出生之间的男性和女性出生之间的原始 C 值也相似(P=0.159)和 AGA 出生(P=0.060),而 RNU44 和 RNU48 的原始 C 值在 SGA 出生时更高(P=0.008 和 0.006),在 SGA 组内的男性出生时更高(P=0.005),而在 AGA 组内的女性出生时更高(P=0.048)。使用 RefFinder 工具测试的所有 106 个样本中,miR-16-5p 和 RNU44 是同等稳定的参考 miRNA。
我们建议 miR-16-5p 和 RNU44 作为与我们研究类似的情况下胎盘样本的合适参考 miRNA。